Pharmabiome analyses in tandem with chemometrics can help trace the provenance of falsified medicines: a proof-of-concept study

A lack of robust analytical approaches limits our ability to investigate the clandestine manufacturing origins of falsified medicines. We conducted a proof-of-concept study to test the feasibility of geolocating the production sites of falsified medicines, based on the identification of site-specific biological and chemo-isotopic features using a combination of environmental DNA metabarcoding, Direct Analysis in Real Time - Mass Spectrometry and Isotope Ratio Mass Spectrometry as profiling techniques. We produced tablets at two distant locations (England vs. Thailand), using controlled manufacturing methods, excipient composition and environmental conditions. Sets of tablets produced at separate locations showed distinct bacterial and eukaryotic diversity, particularly influenced by the incorporation of water used during tableting and the background environmental biosignatures of the production site. Tablets showed corresponding site-specific chemometric profiles, but the factors contributing to the observed chemical differences were unclear. When reference samples of known origin are available, our study suggests that site-specific biological and chemical features can be used in modelling approaches to successfully predict product origin. We developed a new mapping approach to exploit the geographic information within the eukaryotic pharmabiome of the falsifications; based on eDNA-derived species identification and the integration of publicly available species distribution data. In the absence of reference samples of known origin, the application of this workflow to our dataset provided partial clues about the product’s origin, with limitations likely due to taxonomic resolution and the presence of species with wide distribution ranges. Collectively, our research provides experimental support for the development of integrated, multifaceted tools for tracing the origin of falsified medicines, advancing efforts to combat this pervasive but neglected global health proble

Reintroducing a large carnivore in a human dominated landscape: dynamics of an isolated brown bear population over two decades post-reintroduction

Large carnivores have been extirpated from much of their historical range, but conservation efforts have supported their recovery. The brown bear reintroduction in the central Alps represents one such case, yet long-term viability of this population remains uncertain due to its small size, isolation, and increasing conflicts with humans. Using spatial capture-recapture and survival models based on non-invasive genetic data and known bear deaths collected on 222 individuals over 21 years (2003−2023), we assessed spatio-temporal trends in bear density, abundance, survival, and range expansion, accounting for sex and age-specific differences. Results indicate an annual population growth rate of 7.7 %, with a mean density of 1.61 bears/100 km2 in the whole study area in 2023, rising to 2.50 within the area permanently occupied by females. Survival probabilities varied by sex and age, with females showing strong site fidelity and males exhibiting higher dispersal and mortality. Although an ecological barrier hinders eastward expansion and connectivity, the permanent range of females increased by 137 %. Despite growth, the population remains small and vulnerable to stochastic events. Sustained genetic monitoring, improved connectivity, and science-based management will be crucial to balance conservation goals and social concerns, ensuring the persistence of this population in a human-dominated landscap

Crayfish plague and microsporidiosis occurrence in wild populations of the white-clawed crayfish Austropotamobius pallipes complex in Trentino (North-East Italy)

One of the causes of the decline in distribution and abundance of the endangered white-clawed crayfish Austropotamobius pallipes complex throughout Europe is the invasion of alien crayfish, and the associated spread of infectious diseases, primarily the crayfish plague caused by Aphanomyces astaci. Another relevant disease is microsporidiosis (porcelain disease), caused by Astathelohania contejeani and Nosema austropotamobii. Between 2021 and 2024, we conducted a monitoring survey, aimed at mapping the distribution of A. astaci, A. contejeani and N. austropotamobii in wild populations of A. pallipes and in two non-indigenous species in Trentino (North-East Italy). We applied a non-invasive sampling method (cuticular swabs) in 33 populations of A. pallipes, 2 populations of Procambarus clarkii and 4 populations of Faxonius limosus, to investigate the presence of A. astaci. Aphanomyces astaci was detected in eight thriving populations of A. pallipes, and the presence of a low virulence genotype (genotype group A) was confirmed in one of them. Aphanomyces astaci was detected in one of the two populations of P. clarkii, as well as in one of the four populations of F. limosus. No mortality outbreaks in populations of A. pallipes were recorded in Trentino during the study period. Specimens with macroscopic signs of porcelain disease were found in thirteen populations of A. pallipes, abdominal muscle tissues were collected and subjected to molecular evaluation; microsporidiosis was detected in all the tested crayfish: seven populations were infected only by A. contejeani, and in six populations both microsporidia were detected. Crayfish from one population of P. clarkii and one of F. limosus were tested for microsporidiosis as well, but the results were negative. Based on the presence of chronically infected but thriving populations across the Trentino territory, we provide suggestions for the management of A. pallipes populations in conservation actions which require the movement of specimens (translocations, rearing in captivity

Early postharvest application of LED lighting enhances strawberry quality during storage

Strawberry (Fragaria × ananassa Duch.) perishability challenges postharvest quality retention. Traditional postharvest methods have limitations, and early harvesting to extend shelf-life often compromises sensory attributes. This study investigated whether 24 h or 48 h treatments performed at the beginning of postharvest with Red, Blue, Far-Red, or UV-A light-emitting diode (LED) light could improve 'Elsanta' strawberry quality during subsequent 7-day storage at suboptimal temperature (5 °C). Fruit quality attributes, including firmness, weight loss, soluble solids content (SSC), titratable acidity (TA), anthocyanins, and volatile organic compounds (VOCs), were assessed. While LED treatments did not significantly affect firmness, they generally increased weight loss compared to dark controls (particularly after 48 h Blue/Red/Far-Red exposures). Light effects on SSC were complex and duration-dependent. Notably, all LED treatments significantly enhanced total anthocyanin content (especially after 48 h exposure), although instrumental color measurements remained unchanged. Compared to dark controls Blue and Red light (especially after 48 h) significantly increased the concentration of key aroma-related VOCs, for the most esters. In conclusion a brief, early postharvest LED exposure, particularly using Blue and Red light, can trigger lasting beneficial effects, enhancing nutritionally relevant anthocyanins and aroma-defining VOCs during storage at 5 °C. This suggests an 'early signal' mechanism and offers a potentially practical strategy to improve strawberry quality, mitigating negative impacts of commercial early harvesting practices and potentially benefiting other non-climacteric fruit

Impact of oxygen supplementation during fermentation on yeast gene expression and thiol release in wine

Traditionally, oxygen supplementation has been more strongly associated with yeast vitality and sugar consumption kinetics than with aroma metabolism. This study investigated the effects of oxygen exposure during the exponential growth phase of yeast cells on the polyfunctional thiol profile of wine. Using quantitative real-time RT-PCR, the expression of five genes related to amino acid and peptide uptake, as well as two genes associated with β-lyase activity, was analyzed. UPLC/MS-MS was employed to quantify thiol precursors and volatile thiols in must and wine. The results revealed that oxygen upregulated the expression of genes involved in amino acid and peptide uptake, including GAP1, OPT1, OPT2, PTR2, DAL5, and FOT3, some of which are known thiol precursor permeases. Overexpression of those genes in oxygenated trials was correlated with a reduction in thiol precursor residues in wines, particularly GSH-3MH, which was depleted before entry into the stationary phase. Despite increased gene expression, the concentration of GSH-4MMP was not affected and remained constant during the growth phase. Additionally, the expression of the gene responsible for thiol-releasing β-lyases in yeasts, IRC7, was positively correlated with 4MMP concentration in wines when influenced by oxygen treatment during the initial 72 h of alcoholic fermentation. However, oxygen supplementation was associated with STR3 downregulation, which may have contributed to the lower 3 MH release, despite the increased precursor intak

Viscoelastic characterisation of high protein ice cream: predicting tactile sensory properties via time–concentration superposition and large amplitude oscillatory shear (LAOS) rheology

In the present work we report on the exploration of time concentration superposition principle (TCS) and non-linear dynamic rheology (LAOS) as useful instrumental tools for predicting tactile sensory modalities of ice cream at serving temperature (−14 °C). Three common tactile sensory properties of ice cream i.e., resistance to scooping (scoopability), creaminess and gumminess were assessed in high protein formulations differing in their protein to fat ratio (φP/F = 0.9 to 4) and protein source (milk protein concentrate (MPC) vs whey protein isolate-sodium caseinate (WPICAS) 1:1 blend). The complex viscosity – angular frequency data obeyed the TCS principle with the calculated shift factors reflecting effectively the compositional profile of ice creams i.e., ac ∝ φP/F1.16 and φP/F2.23, bc ∝ φP/F−1.27 and φP/F−1.75 for MPC and WPICAS fortified systems. LAOS assessment revealed a clear impact of protein type and φP/F on the shearing deformation of ice creams. MPC fortification and decrease in the φP/F enhanced the shear flowing ability of the ice creams. In all cases, the onset of shear stiffening and thickening behaviour was observed at shear stresses below the flow point, which indicates gel-like or colloid glass-like structures. According to partial least squares regression analysis, the TCS parameters (ac and bc), damping factor (tanδ) and the shear strain (γf) and elastic modulus (log ) at flow point were determined as the most important parameters predicting tactile sensory modalities on large deformation (spooning) such as scoopability, creaminess and gummines

Health, microbiota, and water quality analysis: a case study in a commercial recirculating aquaculture system for yellowtail kingfish (Seriola lalandi)

This study describes the temporal and spatial dynamics of microbiota composition, water quality parameters and fish health status in a commercial Recirculating Aquaculture System (RAS) for yellowtail kingfish (Seriola lalandi) spanning from April to November 2021. Water quality was effectively controlled, with key parameters such as CO2, pH, temperature, nitrogen compounds and hydrogen sulfide (H2S) consistently maintained within safety thresholds reported in the literature. Microbiota composition was analyzed in various fish tissues and RAS components using 16S rRNA gene amplicon sequencing. The predominant genera in RAS samples (tank water, tank wall biofilm, biofilter biofilm, and denitrification unit) included Erythrobacter, Glaciecola, and Nitrospira. In contrast, fish tissue samples (gills, intestine, and skin) were mainly colonized by Aliivibrio, Pseudomonas, and an uncultured genus from the Mycoplasmataceae family. Cluster analysis revealed similarities in bacterial composition between tank water and external fish tissues (skin and gills), with variations over time corresponding to changes in water parameters. Digital PCR assays were developed to target specific fish pathogens such as Vibrio harveyi, Vibrio anguillarum, and Photobacterium damselae piscicida, revealing low or undetectable pathogen levels throughout most of the monitoring period. In tank water, these species showed the highest concentration in August and September. A Multiparametric Semi-quantitative Scoring System (MSSS) was developed for histological examination of gills, intestine, liver, spleen, and kidney. Increased inflammation coincided with higher pathogen presence in the water. However, no clinical signs of vibriosis or pasteurellosis were detected, indicating overall good fish health and a robust immune system. This integrated approach - combining innovative pathogen-specific digital PCR assays, MSSS histological scoring and microbiota profiling offers a valuable framework for monitoring fish health and optimizing production quality in kingfish farming within RAS environments

Exploring regional inequities in food safety practices and food security in Italy: a cross-sectional study

Objective: This study examined regional disparities in food security and food safety knowledge and behavior among Italian adults. Methods: Between January and June 2024, we conducted a cross-sectional anonymous online survey targeting Italian residents aged ≥18. The sample size was calculated a priori assuming a 50% prevalence, 95% confidence, and 3% margin of error, yielding a target of 1067 participants. Validated Italian versions of the Food Security Survey Module (It-FSSM) and the Food Safety Knowledge and Behavior Questionnaire (It-FSKB) were employed to assess participants' knowledge and behaviors. Sociodemographic data, including age, sex, body mass index (BMI), educational level, physical activity, smoking habits, and food apps usage, were collected. Multinomial logistic regression-adjusted for age, sex, BMI, and educational level-was used to evaluate regional differences. Results: Among 1752 participants (70.4% women; mean age: 36.01 ± 13.84 y), those in the South area and Islands were significantly less likely to report high food safety knowledge (relative risk ratio [RRR] = 0.66; 95% confidence interval [CI]: 0.54-0.82; P = 0.000) and high food safety behaviors (RRR = 0.64; 95% CI: 0.52-0.79; P < 0.001), and more likely to experience moderate food insecurity (RRR = 1.64; 95% CI: 1.00-2.69; P = 0.048) compared to participants to the North. Participants in the Center were over twice likely than those in the North to report high food security versus very low (RRR = 2.72; 95% CI: 1.15-6.43; P = 0.023) and were also 30% less likely to use food delivery apps rarely rather than not at all (RRR = 0.70; 95% CI: 0.50-0.97; P = 0.034). Conclusions: This study highlights significant regional disparities, with the South area and Islands facing the greatest challenges. These findings provide evidence to guide targeted public health interventions and policies promoting food safety and security across Italy

Cryosphere and lithology influence the hydrological gradients of high elevation Alpine catchments

In high-elevation systems influenced by receding cryosphere, geomorphology and lithology can strongly influence the hydrology of river networks. During summer 2022–2023, we studied the water temperature, δ18O, pH, major ions, and trace element concentrations at two headwater catchments in the Eastern Italian Alps. We investigated the main streams at the spring and below the confluences with tributaries from glaciers, intact and relict rock glaciers, young moraines, and till deposits. In the non-glacierized catchment (6.3 km2), water temperature increased from 1.6 °C at the intact rock glacier spring to 7.3 ± 1.5 °C at the catchment outlet, despite the inputs from till and rock glacier springs with <3.0 °C waters. In the glacierized catchment (3.7 km2), the proglacial reaches had a water temperature of 6.9 ± 2.6 °C and the inputs from cold rock glacier springs decreased the water temperatures by 2–4 °C along the stream. Due to predisposing lithology, at the glacierized catchment the concentrations of trace elements such as Ni, Al, Mn, Zn, Y, and Li were high along the entire river network except in till and the relict rock glacier springs, which are not influenced by the cryosphere. For both catchment outlets, end-member mixing models estimated 60–65 % contribution from rock glaciers to stream runoff. In both river systems, meltwater from snow and ice was the dominant runoff component, with rainwater accounting for 20–30 % of runoff in the non-glacierized catchment and for <10 % in the glacierized on

Powerful flowers: public perception of grassland aesthetics is strongly related to management and biodiversity

Temperate grasslands provide various cultural ecosystem services that are appreciated in diverse ways. Capturing these diverse appreciations requires different methodological approaches, such as questionnaire surveys and social media analyses. In this study, we combined the potential of both approaches to capture two aspects of what people appreciate in Swiss agricultural grasslands, i.e., the aesthetic quality of differently managed plant communities and the objects frequently found in grassland-based social media images. The two complementary approaches showed that people preferred colourful flower- and species-rich grasslands over grass-dominated and fertilised swards. This appreciation can be predicted from automatic image analysis. Social media analysis highlighted that people mainly photographed flowers, followed by livestock and/or wildlife, but this depended also on the social media platform accessed. In addition, we found differences between (i) conservationists and agricultural professionals in the aesthetic appreciation of the plant communities, as well as between (ii) common visitors and naturalists in their social media content. In conclusion, people's appreciation was clearly related to the intensity of grassland management and the level of biodiversity, with a preference for extensively managed grasslands with diverse flowers and wildlife. Our results suggest that extensive management and ecological restoration can be used to increase cultural grassland ecosystem services by enhancing the richness of forbs, flowers and other attractive wildlife. Thus, targeted management can maintain and enhance the attractiveness of grassland landscapes and subsequently support health benefits, opportunities for recreation, agrotourism as well as culture and heritag